Master Degree / Yüksek Lisans Tezleri

Permanent URI for this collectionhttps://hdl.handle.net/11147/3008

Browse

Filters

20161

Settings

Institution Author: search.filters.institutionAuthor.Güler, SeminayPublisher: search.filters.publisher.Izmir Institute of Technology

Search Results

Now showing 1 - 1 of 1
  • Master Thesis
    Nucleofection Effects on Human T Cells as a Non-Viral Transfection Technique
    (Izmir Institute of Technology, 2016) Güler, Seminay; Nalbant Aldanmaz, Ayten; Nalbant Aldanmaz, Ayten; 04.03. Department of Molecular Biology and Genetics; 04. Faculty of Science; 01. Izmir Institute of Technology
    In times of quickly developing ways of immunological approaches applying on cells, nucleic acid delivery becomes a fundamental process. The ways of delivering nucleic acids mainly affects the consequence of immunological response. The nucleofection technology which is a modified form of electroporation is reliable method for transfection of hard to transfect cells. In the current study, we compared naive CD4 T cells and Jurkat cells in terms of nucleofection efficiency and cell viability. Naive CD4 T cells and Jurkat cells were transfected with pmax GFP at different time points. Out of 5 different nucleofection programs, one for naive CD4 T cells was identified. Jurkat cells were also transfected with one nucleofection program. Flow cytometric analyses of nucleofected naive CD4 T cells and Jurkat cells indicated that transfection efficiency significantly increased at 48h post transfection. We also differentiated naive CD4 T cells into T helper 17 cell phenotype by using certain stimulants and we activated Jurkat cells by using PMA and Ionomycin. We found that GFP was efficiently taken up by both T helper 17 cells and activated Jurkat cells. Therefore, nucleofection significantly holds important place in transfecting of hard to transfect cells including primary human CD4 T cells.